Semi-synthetic glycopeptides with antibiotic activity

ABSTRACT

Semi-synthetic glycopeptides having antibacterial activity are based on modifications of the eremomycin, A82846B, vancomycin, teicoplanin, and A-40,926 scaffolds, in particular, acylation of the sugar moieties on these scaffolds with certain acyl groups; and/or conversion of an acid moiety on the macrocyclic ring of these scaffolds to certain substituted amides; or having a combination of an alkylation modification of the amino substituent on the amino-substituted sugar moiety on these scaffolds with certain alkyl groups or acylation modification of the amino substituent on the amino-substituted sugar moiety on this scaffold with certain alkyl groups, and conversion of the acid moiety on the macrocyclic ring of this scaffolds to certain substituted amides. Also provided are methods for the synthesis of the compounds, pharmaceutical compositions containing the compounds, and methods of use of the compounds for the treatment and/or prophylaxis of diseases, especially bacterial infections.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 60/657,297, filed Feb. 28, 2005, titled SEMI-SYNTHETIC GLYCOPEPTIDESWITH ANTIBIOTIC ACTIVITY, the disclosure of which is incorporated hereinby reference in its entirety and for all purposes.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to novel semi-synthetic glycopeptides havingantibacterial activity, to pharmaceutical compositions comprising thesecompounds, and to a medical method of treatment.

2. Description of Related Art

The emergence of drug resistant bacterial strains has highlighted theneed for synthesizing and identifying antibiotics with improvedactivity. Naturally occurring and semi-synthetic glycopeptideantibiotics used to combat bacteria infections include compounds such aseremomycin (structure A, X═H), A82846B (structure A, X═Cl), vancomycin,teicoplanin, and A-40,926, having the scaffolds A, B, C and D, belowrespectively:

These compounds are used to treat and prevent bacterial infection, butas with other antibacterial agents, bacterial strains having resistanceor insufficient susceptibility to these compounds have been identified,and these compounds have been found to have limited effect againstcertain bacterial infections caused by glycopeptide resistantenterococci. Therefore, there is a continuing need to identify newderivative compounds which possess improved antibacterial activity,which have less potential for developing resistance, which possessimproved effectiveness against bacterial infections that resisttreatment with currently available antibiotics, or which possessunexpected selectivity against target microorganisms.

SUMMARY OF THE INVENTION

To achieve the foregoing, the present invention provides novelsemi-synthetic glycopeptides that have antibacterial activity. Thesemi-synthetic glycopeptides of the invention are based on modificationsof the eremomycin, A82846B, vancomycin, teicoplanin, and A-40,926scaffolds, in particular, acylation of the amino substituent on theamino-substituted sugar moiety on these scaffolds with certain acylgroups, in particular amino acids or derivatives thereof; and/orconversion of the acid moiety on the macrocyclic ring of these scaffoldsto certain substituted amides; or having a combination of an alkylationmodification of the amino substituent on the amino-substituted sugarmoiety on these scaffolds with certain alkyl groups or acylationmodification of the amino substituent on the amino-substituted sugarmoiety on this scaffold with certain alkyl groups, including β-aminoacids or derivatives thereof, and conversion of the acid moiety on themacrocyclic ring of this scaffolds to certain substituted amides. Alsoprovided are methods for synthesis of the compounds, pharmaceuticalcompositions containing the compounds, and methods of use of thecompounds for the treatment and/or prophylaxis of diseases, especiallybacterial infections.

In specific embodiments of the invention, the eremomycin, A82846B,vancomycin, teicoplanin, and A-40,926 scaffolds are modified to make acompound having a formula selected from the group consisting of:

wherein,

-   -   R₁ is C(═O)CR₇R_(7a)NR₈R_(8a), wherein,        R₇ and R_(7a) are independently hydrogen, the side chain of a        naturally occurring or non-naturally occurring amino acid,        alkyl, or alkyl substituted with one or more substituents        selected from the group consisting of halogen, hydroxy, alkoxy,        alkoxyalkoxy, carboxyl, carboxyl ester, —C(═O)NR₈R_(8a),        —NR₈R_(8a), aryl, substituted aryl, heteroaryl, substituted        heteroaryl, mercapto, or thioalkoxy, or R₇ and R_(7a) together        with the atom to which they are attached form a cycloalkyl ring        which optionally contains a heteroatom selected from the group        consisting of optionally substituted O, N, and S;        R₈ and R_(8a) are independently selected from the group        consisting of hydrogen and unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl,        arylalkyl, alkylaryl, and heteroaryl, said aryl, alkylaryl,        arylalkyl or heteroaryl group optionally containing one or more        optionally substituted aryl, heteroaryl, or condensed rings, or        R₈ and R_(8a) together with the atom to which they are attached        form a cycloalkyl ring which optionally contains a heteroatom        selected from the group consisting of optionally substituted O,        N, and S;    -   R_(1A) is selected from the group consisting of H, CHR₅R_(5a),        and C(═O)R₆, wherein,        R₅ and R_(5a) are independently selected from the group        consisting of hydrogen and unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl,        arylalkyl, alkylaryl, and heteroaryl, said aryl, alkylaryl,        arylalkyl or heteroaryl group optionally containing one or more        optionally substituted aryl, heteroaryl, or condensed rings, or        R₅ and R_(5a) together with the atom to which they are attached        form a cycloalkyl ring which optionally contains a heteroatom        selected from the group consisting of optionally substituted O,        N, and S, and        R₆ is selected from the group consisting of unsubstituted or        substituted alkyl, alkenyl, cycloalkyl, cycloalkenyl,        heterocycloalkyl, aryl, arylalkyl, alkylaryl, and heteroaryl        containing a heteroatom selected from the group consisting of        optionally substituted O, N, and S, said aryl, alkylaryl,        arylalkyl or heteroaryl group optionally containing one or more        optionally substituted aryl, heteroaryl, or condensed rings;    -   R₂ is selected from the group consisting of,        -   (1) OH,        -   (2) 1-adamantanamino,        -   (3) 2-adamantanamino,        -   (4) 3-amino-1-adamantanamino,        -   (5) 1-amino-3-adamantanamino,        -   (6) 3-loweralkylamino-1-adamantanamino,        -   (7) 1-loweralkylamino-3-adamantanamino,        -   (8) amino,        -   (9) NR₉R_(9a) wherein R₉ and R_(9a) are independently            selected from the group consisting of hydrogen, loweralkyl            or substituted loweralkyl, or        -    R₉ and R_(9a) together with the atom to which they are            attached form a 3-10 membered heterocycloalkyl ring, which            may optionally be substituted with one or more substituents            independently selected from the group consisting of            -   (a) halogen,            -   (b) hydroxy,            -   (c) C₁-C₃-alkoxy,            -   (d) C₁-C₃-alkoxy-C₁-C₃-alkoxy,            -   (e) oxo,            -   (f) C₁-C₃-alkyl,            -   (g) halo-C₁-C₃-alkyl, and            -   (h) C₁-C₃-alkoxy-C₁-C₃-alkyl;    -   R_(2A) is selected from the group consisting of        -   (1) 1-adamantanamino,        -   (2) 2-adamantanamino,        -   (3) 3-amino-1-adamantanamino,        -   (4) 1-amino-3-adamantanamino,        -   (5) 3-loweralkylamino-1-adamantanamino,        -   (6) 1-loweralkylamino-3-adamantanamino; and    -   R₃ is selected from the group consisting of hydrogen and        aminoloweralkyl, wherein the aminoloweralkyl amino group is        further substituted with unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, aryl, arylalkyl, alkylaryl,        alkoxy, aryloxy, substituted alkoxy, and substituted aryloxy;        or a pharmaceutically acceptable salt, ester, solvate,        stereoisomer, tautomer or prodrug thereof.

The present invention also provides pharmaceutical compositions whichcomprise a therapeutically effective amount of a compound as definedabove in combination with a pharmaceutically acceptable carrier.

The invention further relates to methods of treating bacterialinfections in a host mammal in need of such treatment comprisingadministering to a mammal in need of such treatment a therapeuticallyeffective amount of a compound of the invention as defined above.

In a further aspect of the present invention are provided processes forthe preparation of semi-synthetic glycopeptides defined above.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The materials and associated techniques and apparatuses of the presentinvention will now be described with reference to several embodiments.Important properties and characteristics of the described embodimentsare illustrated in the structures in the text. While the invention willbe described in conjunction with these embodiments, it should beunderstood that the invention it is not intended to be limited to theseembodiments. On the contrary, it is intended to cover alternatives,modifications, and equivalents as may be included within the spirit andscope of the invention as defined by the appended claims. In thefollowing description, numerous specific details are set forth in orderto provide a thorough understanding of the present invention. Thepresent invention may be practiced without some or all of these specificdetails. In other instances, well known process operations have not beendescribed in detail in order not to unnecessarily obscure the presentinvention.

Introduction

The present invention provides novel semi-synthetic glycopeptides thathave antibacterial activity. The semi-synthetic glycopeptides of theinvention are based on modifications of the eremomycin, A82846B,vancomycin, teicoplanin, and A-40,926 scaffolds, in particular,acylation of the amino substituent on the amino-substituted sugar moietyon these scaffolds with certain acyl groups; and conversion of the acidmoiety on the macrocyclic ring of these scaffolds to certain substitutedamides. Also provided are methods for synthesis of the compounds,pharmaceutical compositions containing the compounds, and methods of useof the compounds for the treatment and/or prophylaxis of diseases,especially bacterial infections.

Compounds of the Invention

In specific embodiments of the invention, the eremomycin, A82846B,vancomycin, teicoplanin, and A-40,926 scaffolds are modified to make acompound having a formula selected from the group consisting of:

wherein,

-   -   R₁ is C(═O)CR₇R_(7a)NR₈R_(8a), wherein,        R₇ and R_(7a) are independently hydrogen, the side chain of a        naturally occurring or non-naturally occurring amino acid,        alkyl, or alkyl substituted with one or more substituents        selected from the group consisting of halogen, hydroxy, alkoxy,        alkoxyalkoxy, carboxyl, carboxyl ester, —C(═O)NR₈R_(8a),        —NR₈R_(8a), aryl, substituted aryl, heteroaryl, substituted        heteroaryl, mercapto, or thioalkoxy, or R₇ and R_(7a) together        with the atom to which they are attached form a cycloalkyl ring        which optionally contains a heteroatom selected from the group        consisting of optionally substituted O, N, and S;        R₈ and R_(8a) are independently selected from the group        consisting of hydrogen and unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl,        arylalkyl, alkylaryl, and heteroaryl, said aryl, alkylaryl,        arylalkyl or heteroaryl group optionally containing one or more        optionally substituted aryl, heteroaryl, or condensed rings, or        R₈ and R_(8a) together with the atom to which they are attached        form a cycloalkyl ring which optionally contains a heteroatom        selected from the group consisting of optionally substituted O,        N, and S;    -   R_(1A) is selected from the group consisting of H, CHR₅R_(5a),        and C(═O)R₆, wherein,        R₅ and R_(5a) are independently selected from the group        consisting of hydrogen and unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl,        arylalkyl, alkylaryl, and heteroaryl, said aryl, alkylaryl,        arylalkyl or heteroaryl group optionally containing one or more        optionally substituted aryl, heteroaryl, or condensed rings, or        R₅ and R_(5a) together with the atom to which they are attached        form a cycloalkyl ring which optionally contains a heteroatom        selected from the group consisting of optionally substituted O,        N, and S, and        R₆ is selected from the group consisting of unsubstituted or        substituted alkyl, alkenyl, cycloalkyl, cycloalkenyl,        heterocycloalkyl, aryl, arylalkyl, alkylaryl, and heteroaryl        said aryl, alkylaryl, arylalkyl or heteroaryl group optionally        containing one or more optionally substituted aryl, heteroaryl,        or condensed rings;    -   R₂ is selected from the group consisting of,        -   (1) OH,        -   (2) 1-adamantanamino,        -   (3) 2-adamantanamino,        -   (4) 3-amino-1-adamantanamino,        -   (5) 1-amino-3-adamantanamino,        -   (6) 3-loweralkylamino-1-adamantanamino,        -   (7) 1-loweralkylamino-3-adamantanamino,        -   (8) amino,        -   (9) NR₉R_(9a) wherein R₉ and R_(9a) are independently            selected from the group consisting of hydrogen, loweralkyl            or substituted loweralkyl, or        -    R₉ and R_(9a) together with the atom to which they are            attached form a 3-10 membered heterocycloalkyl ring, which            may optionally be substituted with one or more substituents            independently selected from the group consisting of            -   (a) halogen,            -   (b) hydroxy,            -   (c) C₁-C₃-alkoxy,            -   (d) C₁-C₃-alkoxy-C₁-C₃-alkoxy,            -   (e) oxo,            -   (f) C₁-C₃-alkyl,            -   (g) halo-C₁-C₃-alkyl, and            -   (h) C₁-C₃-alkoxy-C₁-C₃-alkyl;    -   R_(2A) is selected from the group consisting of        -   (1) 1-adamantanamino,        -   (2) 2-adamantanamino,        -   (3) 3-amino-1-adamantanamino,        -   (4) 1-amino-3-adamantanamino,        -   (5) 3-loweralkylamino-1-adamantanamino,        -   (6) 1-loweralkylamino-3-adamantanamino; and    -   R₃ is selected from the group consisting of hydrogen and        aminoloweralkyl, wherein the aminoloweralkyl amino group is        further substituted with unsubstituted or substituted alkyl,        alkenyl, cycloalkyl, cycloalkenyl, aryl, arylalkyl, alkylaryl,        alkoxy, aryloxy, substituted alkoxy, and substituted aryloxy;        or a pharmaceutically acceptable salt, ester, solvate,        stereoisomer, tautomer or prodrug thereof.

According to specific embodiments of the invention, the varioussubstituents may be as follows:

Within R_(1A):

R₅ may be hydrogen and R_(5a) may be selected from the group consistingof unsubstituted or substituted alkyl, alkenyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, aryl, arylalkyl, alkylaryl, andheteroaryl, said aryl, alkylaryl, arylalkyl or heteroaryl groupoptionally containing one or more optionally substituted aryl,heteroaryl, or condensed rings, or R₅ and R_(5a) together with the atomto which they are attached form a cycloalkyl ring which optionallycontains a heteroatom selected from the group consisting of optionallysubstituted O, N, and S.

R₆ may be β-amino acid analog. Such a group will include a—CH₂CHNH—portion. For example, R₆ may be CH₂C(R₇)(R_(7a))(NR₈R_(8a))wherein R₇, R_(7a), R₈, and R_(8a) are previously defined or —CR₇R_(7a)together with NR₈R_(8a) form a pyrrolidine ring.

Within R₁, the C(═O)CR₇R_(7a)NR₈R_(8a) may be an amino acid moiety, suchthat R₇, R₈ and R_(8a) are each H and R_(7a) is one of H, CH₃, CH(CH₃)₂,CH₂CH(CH₃)₂, CH(CH₃)CH₂CH₃, (CH₂)₄NH₂, CH₂OH, CH(OH)CH₃, CH₂COOH,(CH₂)₂COOH, CH₂C(═O)NH₂, (CH₂)₂C(═O)NH₂, CH₂SH, (CH₂)₂SCH₃,(CH₂)₃NHC(═NH)NH₂, CH₂C₆H₅, CH₂C₆H₄OH, CH₂(4-imidazoyl) orCH₂(3-indolyl), or —CR₇R_(7a) together with NR₈R_(8a) form a pyrrolidinering.

Alternatively, R₇ may be H and R_(7a) may be selected from the groupconsisting of

-   -   (1) hydrogen,    -   (2) C₁-C₁₂-alkyl, and    -   (3) C₁-C₁₂-alkyl substituted with one or more substituents        selected from the group consisting of        -   (a) halogen,        -   (b) hydroxy,        -   (c) C₁-C₃-alkoxy,        -   (d) C₁-C₃-alkoxy-C₁-C₃-alkoxy,        -   (e) —CO₂R₅ wherein R₅ is hydrogen, loweralkyl or substituted            loweralkyl,        -   (f) —C(═O)NR₉R_(9a),        -   (g) amino, and        -   (h)—NR₉R_(9a), or        -    R₉ and R_(9a) together with the atom to which they are            attached form a 3-10 membered heterocycloalkyl ring            optionally substituted with one or more substituents            independently selected from the group consisting of            -   (i) halogen.            -   (ii) hydroxy,            -   (iii) C₁-C₃-alkoxy,            -   (iv) C₁-C₃-alkoxy-C₁-C₃-alkoxy,            -   (v) oxo,            -   (vi) C₁-C₃-alkyl,            -   (vii) halo-C₁-C₃-alkyl, and            -   (viii) C₁-C₃-alkoxy-C₁-C₃-alkyl,        -   (i) aryl,        -   (j) substituted aryl,        -   (k) heteroaryl,        -   (l) substituted heteroaryl,        -   (m) mercapto, and        -   (n) C₁-C₃-thioalkoxy.

In addition, R₈ and R_(8a) may be independently selected from the groupconsisting of,

-   -   (1) hydrogen,    -   (2) C₁-C₁₂-alkyl,    -   (3) C₂-C₁₂-alkyl substituted with one or more substituents        selected from the group consisting of        -   (a) halogen,        -   (b) hydroxy,        -   (c) C₁-C₃-alkoxy,        -   (d) C₁-C₃-alkoxy-C₁-C₃-alkoxy,        -   (e) amino, and        -   (f) C₁-C₃-alkylamino,    -   (4) C₁-C₁₂-alkyl substituted with aryl,    -   (5) C₁-C₁₂-alkyl substituted with substituted aryl,    -   (6) C₁-C₁₂-alkyl substituted with heteroaryl, and    -   (7) C₁-C₁₂-alkyl substituted with substituted heteroaryl; or        R₈ and R_(8a) together with the atom to which they are attached        form a C₃-C₇-heterocycloalkyl ring which, when the ring is a 5-        to 7-membered ring, optionally contains a hetero function        selected from the group consisting of —O—, —NH,        —N(C₁-C₆-alkyl-)-, —N(aryl)-, —N(aryl-C₁-C₆-alkyl-)-, —N        (substituted-aryl-C₁-C₆-alkyl-)-, —N(heteroaryl)-,        —N(heteroaryl-C₁-C₆-alkyl-)-,        —N(substituted-heteroaryl-C₁-C₆-alkyl-)-, and —S— or S(═O)_(n)—        wherein n is 1 or 2.

In a specific embodiment, the compound may be one of N′-p-BuBnHNCH₂COeremomycin, N′-stilbenylHNCH₂CO eremomycin, N′-p-C₈H₁₇OBnHNCH₂COvancomycin, N′-p-C₆H₁₇OBnHNCH(CH₃)CO vancomycin and 2-adamantanaminoeremomycin.

Definitions

Unless otherwise noted, terminology used herein should be given itsnormal meaning as understood by one of skill in the art. In order tofacilitate understanding of the present invention, a number of definedterms are used herein to designate particular elements of the presentinvention. When so used, the following meanings are intended:

The term “alkyl” as used herein refers to saturated, straight- orbranched-chain hydrocarbon radicals derived from a hydrocarbon moietycontaining between one and twenty carbon atoms by removal of a singlehydrogen atom.

The term “alkenyl” as used herein refers to unsaturated, straight- orbranched-chain hydrocarbon radicals derived from a hydrocarbon moietycontaining between two and twenty carbon atoms by removal of a singlehydrogen atom.

The term “cycloalkyl” as used herein refers to a monovalent groupderived from a monocyclic or bicyclic saturated carbocyclic ringcompound containing between three and twenty carbon atoms by removal ofa single hydrogen atom.

The term “cycloalkenyl” as used herein refers to a monovalent groupderived from a monocyclic or bicyclic unsaturated carbocyclic ringcompound containing between three and twenty carbon atoms by removal ofa single hydrogen atom.

The terms “C₁-C₃-alkyl”, “C₁-C₆-alkyl”, and “C₁-C₁₂-alkyl” as usedherein refer to saturated, straight- or branched-chain hydrocarbonradicals derived from a hydrocarbon moiety containing between one andthree, one and six, and one and twelve carbon atoms, respectively, byremoval of a single hydrogen atom. Examples of C₁-C₃-alkyl radicalsinclude methyl, ethyl, propyl and isopropyl. Examples of C₁-C₆-alkylradicals include, but not limited to, methyl, ethyl, propyl, isopropyl,n-butyl, tert-butyl, neopentyl and n-hexyl. Examples of C₁-C₁₂-alkylradicals include, but not limited to, methyl, ethyl, propyl, isopropyl,n-butyl, tert-butyl, neopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl,n-decyl, n-undecyl and n-docecyl.

The term substituted loweralkyl as used herein refers to C₁-C₁₂-alkylsubstituted by one, two or three groups consisting of halogen, alkoxy,amino, alkylamino, dialkylamino, hydroxy, aryl, heteroaryl, alkene oralkyne groups.

The term “C₃-C₁₂-cycloalkyl” denotes a monovalent group derived from amonocyclic or bicyclic saturated carbocyclic ring compound by removal ofa single hydrogen atom. Examples include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl.

The terms “C₁-C₃-alkoxy”, “C₁-C₆-alkoxy” as used herein refers to theC₁-C₃-alkyl group and C₁-C₆-alkyl group, as previously defined, attachedto the parent molecular moiety through an oxygen atom. Examples ofC₁-C₆-alkoxy radicals include, but not limited to, methoxy, ethoxy,propoxy, isopropoxy, n-butoxy, tert-butoxy, neopentoxyl and n-hexoxy.

The term “oxo” denotes a group wherein two hydrogen atoms on a singlecarbon atom in an alkyl group as defined above are replaced with asingle oxygen atom (i.e., a carbonyl group).

The term “aryl” as used herein refers to a mono- or bicyclic carbocylicring system having one or two aromatic rings including, but not limitedto, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the likeand can be un-substituted or substituted (including bicyclic arylgroups) with one, two or three substituents independently selected fromloweralkyl, substituted loweralkyl, haloalkyl, C₁-C₁₂-alkoxy,thioalkoxy, C₁-C₁₂-thioalkoxy, aryloxy, amino, alkylamino, dialkylamino,acylamino, cyano, hydroxy, halogen, mercapto, nitro, carboxaldehyde,carboxy, alkoxycarbonyl and carboxamide. In addition, substituted arylgroups include tetrafluorophenyl and pentafluorophenyl.

The term “arylalkyl” as used herein refers to an aryl group as definedabove attached to the parent molecular moiety through an alkyl groupwherein the alkyl group is of one to twelve carbon atoms.

The term “alkylaryl” as used herein refers to an alky group as definedabove attached to the parent molecular moiety through an aryl group.

The term “halo” and “halogen” as used herein refer to an atom selectedfrom fluorine, chlorine, bromine and iodine.

The term “alkylamino” refers to a group having the structure —NHR′wherein R′ is alkyl, as previously defined. Examples of alkylaminoinclude methylamino, ethylamino, iso-propylamino, and the like.

The term “loweralkylamino” as used herein refers to C₁-C₆-alkyl groups,as previously defined, attached to the parent molecular moiety through anitrogen atom. Examples of C₁-C₃-alkylamino include, but are not limitedto methylamino, dimethylamino, ethylamino, diethylamino, andpropylamino.

The term “dialkylamino” refers to a group having the structure —NHR′R″wherein R′ and R″ are independently selected from alkyl, as previouslydefined. Additionally, R′ and R″ taken together may optionally be—(CH₂)_(k)— where k is an integer of from 2 to 6. Examples ofdialkylamino include dimethylamino, diethylamino, methylpropylamino,piperidino, and the like.

The term “haloalkyl” denotes an alkyl group, as defined above, havingone, two or three halogen atoms attached thereto and is exemplified bysuch group as chloromethyl, bromoethyl, trifluoromethyl, and the like.

The term “alkoxycarbonyl” represents as ester group; i.e. an alkoxygroup, attached to the parent molecular moiety through a carbonyl groupsuch as methoxycarbonyl, ethoxycarbonyl, and the like.

The term “thioalkoxy” refers to an alkyl group previously definedattached to the parent molecular moiety through a sulfur atom.

The term “carboxaldehyde” as used herein refers to a group of formula—CHO.

The term “carboxy” as used herein refers to a group of formula —CO₂H.

The term “carboxamide” as used herein refers to a group of formula—CONHR′R″ wherein R′ and R″ are independently selected from hydrogen,alkyl, or R′ and R″ taken together may optionally be —(CH₂)_(k)— where kis an integer of from 2 to 6.

The term “heteroaryl”, as used herein, refers to a cyclic or bicyclicaromatic radical having from five to ten ring atoms in each ring ofwhich at least one atom of the cyclic or bicyclic ring is selected fromoptionally substituted S, O, and N; zero, one or two ring atoms areadditional heteroatoms independently selected from optionallysubstituted S, O, and N; and the remaining ring atoms are carbon, theradical being joined to the rest of the molecule via any of the ringatoms, such as, for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl,pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl,oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl,naphthyridinyl; and the like.

The term “heterocycloalkyl” as used herein, refers to a non-aromaticpartially unsaturated or fully saturated 3- to 10-membered ring system,which includes single rings of 3 to 8 atoms in size and bi- ortri-cyclic ring systems which may include aromatic six-membered aryl orheteroaryl rings fused to a non-aromatic ring. These heterocycloalkylrings include those having from one to three heteroatoms independentlyselected from oxygen, sulfur and nitrogen, in which the nitrogen andsulfur heteroatoms may optionally be oxidized and the nitrogenheteroatom may optionally be quaternized. Representativeheterocycloalkyl rings include, but not limited to, pyrrolidinyl,pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl,isothiazolidinyl, and tetrahydrofuryl.

The term “heteroarylalkyl” as used herein, refers to a heteroaryl groupas defined above attached to the parent molecular moiety through analkyl group wherein the alkyl group is of one to twelve carbon atoms.

“Protecting group” refers to an easily removable group which is known inthe art to protect a functional group, for example, a hydroxyl, ketoneor amine, against undesirable reaction during synthetic procedures andto be selectively removable. The use of protecting groups is well knownin the art for protecting groups against undesirable reaction duringsynthetic procedure and many such protecting groups are known, cf., forexample, T. H. Greene and P. G. M. Wuts, Protective Groups in OrganicSynthesis, 2nd edition, John Wiley & Sons, New York (1991). Examples ofhydroxy-protecting groups include, but are not limited to,methylthiomethyl, tert-dimethylsilyl, tert-butyldiphenylsilyl, etherssuch as methoxymethyl, and esters including acetyl, benzoyl, and thelike. Examples of ketone protecting groups include, but are not limitedto, ketals, oximes, O-substituted oximes for example O-benzyl oxime,O-phenylthiomethyl oxime, 1-isopropoxycyclohexyl oxime, and the like.Examples of amine protecting groups include, but are not limited to,tert-butoxycarbonyl (Boc) and carbobenzyloxy (Cbz).

The term “amino acid” refers to amino acids having D or Lstereochemistry, and also refers to synthetic, non-natural amino acidshaving side chains other than those found in the 20 common amino acids.Non-natural amino acids are commercially available or may be preparedaccording to U.S. Pat. No. 5,488,131 and references therein. Amino acidsmay be further substituted to contain modifications to their amino,carboxy, or side chain groups. These modifications include the numerousprotecting groups commonly used in peptide synthesis (T. H. Greene andP. G. M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, JohnWiley & Sons, New York, 1991).

The term “substituted aryl” as used herein, refers to an aryl group asdefined herein substituted by independent replacement of one, two orthree of the hydrogen atoms thereon with Cl, Br, F, I, OH, CN,C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, C₁-C₁₂-alkoxy substituted with aryl,C1-C12-alkoxy substituted with substituted aryl, haloalkyl, thioalkyl,amino, alkylamino, dialkylamino, mercapto, nitro, carboxaldehyde,carboxy, alkoxycarbonyl and carboxamide. In addition, any onesubstituent may be an aryl, heteroaryl, or hetercycloalkyl group.

The term “substituted heteroaryl” as used herein, refers to a heteroarylgroup as defined herein substituted by independent replacement of one,two or three of the hydrogen atoms thereon with Cl, Br, F, I, OH, CN,C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, C₁-C₁₂-alkoxy substituted with aryl,haloalkyl, thioalkyl, amino, alkylamino, dialkylamino, mercapto, nitro,carboxaldehyde, carboxy, alkoxycarbonyl and carboxamide. In addition,any one substituent may be an aryl, heteroaryl, or hetercycloalkylgroup.

The term “substituted heterocycloalkyl” as used herein, refers to aheterocycloalkyl group as defined herein substituted by independentreplacement of one, two or three of the hydrogen atoms thereon with Cl,Br, F, I, OH, CN, C₁-C₁₂-alkyl, C₁-C₁₂-alkoxy, C₁-C₁₂-alkoxy substitutedwith aryl, haloalkyl, thioalkyl, amino, alkylamino, dialkylamino,mercapto, nitro, carboxaldehyde, carboxy, alkoxycarbonyl andcarboxamide. In addition, any one substituent may be an aryl,heteroaryl, or hetercycloalkyl group.

The term “adamantanamino” as used herein, refers to a fully saturatedtricyclo [3.3.1.1(3,7)] 10-membered carbon ring system with one or moreamino substituents. Examples include 1-adamantanamino, 2-adamantanamino,3-amino-1-adamantanamino, 1-amino-3-adamantanamino,3-loweralkylamino-1-adamantanamino, and1-loweralkylamino-3-adamantanamino.

The term “stereoisomer” as used herein, refers to either of two forms ofa compound having the same molecular formula and having theirconstituent atoms attached in the same order, but having differentarrangement of their atoms in space about an asymmetric center. Numerousasymmetric centers may exist in the compounds of the present invention.Except where otherwise noted, the present invention contemplates thevarious stereoisomers and mixtures thereof. Accordingly, except whereotherwise noted, it is intended that a mixture of stereo-orientations oran individual isomer of assigned or unassigned orientation may bepresent.

The term “tautomer” as used herein refers to either of the two forms ofa chemical compound that exhibits tautomerism, which is the ability ofcertain chemical compounds to exist as a mixture of two interconvertibleisomers in equilibrium via hydrogen transfer. The keto and enol forms ofcarbonyl compounds are examples of tautomers. They are interconvertiblein the presence of traces of acids and bases via a resonance stabilizedanion, the enolate ion.

The term “pharmaceutically acceptable salt” refers to those salts whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977), incorporated herein byreference. The salts can be prepared in situ during the final isolationand purification of the compounds of the invention, or separately byreacting the free base function with a suitable organic acid. Examplesof pharmaceutically acceptable, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, oxalic acid, maleic acid,tartaric acid, citric acid, succinic acid or malonic acid or by usingother methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

The term “pharmaceutically acceptable ester” refers to esters whichhydrolyze in vivo and include those that break down readily in the humanbody to leave the parent compound or a salt thereof. Suitable estergroups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Representativeexamples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “solvate” as used herein refers to a compound formed bysalvation, the combination of solvent molecules with molecules or ionsof solute composed of a compound according to the present invention. Theterm “pharmaceutically acceptable solvate” refers to those solvateswhich are, within the scope of sound medical judgment, suitable for usein contact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable solvates are well known in the art.

The term “pharmaceutically acceptable prodrugs” refers to those prodrugsof the compounds of the present invention which are, within the scope ofsound medical judgment, suitable for use in contact with the tissues ofhumans and lower animals with undue toxicity, irritation, allergicresponse, and the like, commensurate with a reasonable benefit/riskratio, and effective for their intended use, as well as the zwitterionicforms, where possible, of the compounds of the invention. The term“prodrug” refers to compounds that are rapidly transformed in vivo toyield the parent compound of the above formula, for example byhydrolysis in blood. A thorough discussion is provided in T. Higuchi andV. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S.Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers inDrug Design, American Pharmaceutical Association and Pergamon Press,1987, both of which are incorporated herein by reference.

Synthetic Methods

Synthesis of the compounds of the invention can be broadly summarized asfollows. The compounds of the invention may be made by couplingfunctionalized or unfunctionalized glycopeptides with the appropriateacyl, alkyl and/or amino groups under amide formation conditions. Inparticular, the semi-synthetic glycopeptides of the invention are madeby modifying an eremomycin, A82846B, vancomycin, teicoplanin or A-40,926scaffold, in particular, by acylation of the amino substituent on theamino-substituted sugar moiety on this scaffold with certain acylgroups, in particular amino acids or derivatives thereof; and/orconversion of the acid moiety on the macrocyclic ring of this scaffoldsto certain substituted amides; or having a combination of an alkylationmodification of the amino substituent on the amino-substituted sugarmoiety on this scaffold with certain alkyl groups or acylationmodification of the amino substituent on the amino-substituted sugarmoiety on this scaffold with certain alkyl groups, including β-aminoacids or derivatives thereof, and conversion of the acid moiety on themacrocyclic ring of this scaffolds to certain substituted amides.

In particular, the semi-synthetic glycopeptides of the invention may bemade by modifying one of an eremomycin, A82846B, vancomycin, teicoplaninor A-40,926 scaffold,

by a technique selected from the group consisting of,

-   -   (a) acylation of the amino substituent on the amino-substituted        sugar moiety of the compound with an acyl group having the        structure,        —C(═O)CR₇R_(7a)NR₈R_(8a),    -   (b) conversion of the acid moiety on the macrocyclic ring of the        compound with a substituted amide as defined by R₂, and    -   (c) a combination of (a) and (b)    -   (d) a combination of (b) and acylation of the amino substituent        on the amino-substituted sugar moiety of the compound with an        acyl group having the structure,        —C(═O)R₆,    -   (e) a combination of (b) and alkylation of the amino substituent        on the amino-substituted sugar moiety of the compound with an        alkyl group having the structure,        CHR₅R_(5a),        to form a compound having a formula selected from the group        consisting of:        wherein R₁, R_(1A), R₂, R_(2A), R₃, R₅, R_(5a), R₆, R₇, R_(7a),        R₈, and R_(8a) are as defined herein.

Synthesis of compounds may also involve the use of protecting orblocking groups in order to maximize yields, minimize unwanted sideproducts, or improve the ease purification. Specific examples ofsyntheses for compounds in accordance with the present invention areprovided in the Examples, below.

Pharmaceutical Compositions

Pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), bucally, or as an oral or nasal spray, or a liquid aerosol ordry powder formulation for inhalation.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform may be accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations may also be prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,acetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulations, ear drops, and the like are also contemplatedas being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Compositions of the invention may also be formulated for delivery as aliquid aerosol or inhalable dry powder. Liquid aerosol formulations maybe nebulized predominantly into particle sizes that can be delivered tothe terminal and respiratory bronchioles where bacteria reside inpatients with bronchial infections, such as chronic bronchitis andpneumonia. Pathogenic bacteria are commonly present throughout airwaysdown to bronchi, bronchioli and lung parenchema, particularly interminal and respiratory bronchioles. During exacerbation of infection,bacteria can also be present in alveoli. Liquid aerosol and inhalabledry powder formulations are preferably delivered throughout theendobronchial tree to the terminal bronchioles and eventually to theparenchymal tissue.

Aerosolized formulations of the invention may be delivered using anaerosol forming device, such as a jet, vibrating porous plate orultrasonic nebulizer, preferably selected to allow the formation of aaerosol particles having with a mass medium average diameterpredominantly between 1 to 5μ. Further, the formulation preferably hasbalanced osmolarity ionic strength and chloride concentration, and thesmallest aerosolizable volume able to deliver effective dose of thecompounds of the invention to the site of the infection. Additionally,the aerosolized formulation preferably does not impair negatively thefunctionality of the airways and does not cause undesirable sideeffects.

Aerosolization devices suitable for administration of aerosolformulations of the invention include, for example, jet, vibratingporous plate, ultrasonic nebulizers and energized dry powder inhalers,that are able to nebulize the formulation of the invention into aerosolparticle size predominantly in the size range from 1-5μ. Predominantlyin this application means that at least 70% but preferably more than 90%of all generated aerosol particles are within 1-5μ range. A jetnebulizer works by air pressure to break a liquid solution into aerosoldroplets. Vibrating porous plate nebulizers work by using a sonic vacuumproduced by a rapidly vibrating porous plate to extrude a solventdroplet through a porous plate. An ultrasonic nebulizer works by apiezoelectric crystal that shears a liquid into small aerosol droplets.A variety of suitable devices are available, including, for example,AeroNeb™ and AeroDose™ vibrating porous plate nebulizers (AeroGen, Inc.,Sunnyvale, Calif.), Sidestream® nebulizers (Medic-Aid Ltd., West Sussex,England), Pari LC® and Pari LC Star® jet nebulizers (Pari RespiratoryEquipment, Inc., Richmond, Va.), and Aerosonic™ (DeVilbiss MedizinischeProdukte (Deutschland) GmbH, Heiden, Germany) and UltraAire® (OmronHealthcare, Inc., Vernon Hills, Ill.) ultrasonic nebulizers.

Compounds of the invention may also be formulated for use as topicalpowders and sprays that can contain, in addition to the compounds ofthis invention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

According to the methods of treatment of the present invention,bacterial infections are treated or prevented in a patient such as ahuman or lower mammal by administering to the patient a therapeuticallyeffective amount of a compound of the invention, in such amounts and forsuch time as is necessary to achieve the desired result. By a“therapeutically effective amount” of a compound of the invention ismeant a sufficient amount of the compound to treat bacterial infections,at a reasonable benefit/risk ratio applicable to any medical treatment.It will be understood, however, that the total daily usage of thecompounds and compositions of the present invention will be decided bythe attending physician within the scope of sound medical judgment. Thespecific therapeutically effective dose level for any particular patientwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts.

The total daily dose of the compounds of this invention administered toa human or other mammal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 2000 mg of the compound(s) of this invention per day in singleor multiple doses.

EXAMPLES

The following examples provide details concerning the synthesis,properties and activities and applications of semi-syntheticglycopeptides in accordance with the present invention. It should beunderstood the following is representative only, and that the inventionis not limited by the detail set forth in these examples.

Example 1 Synthesis of N′-Alkylaminoacylated Eremomycin and VancomycinDerivatives

N′-Alkylaminoacylated eremomycin or vancomycin derivatives were preparedby the treatment of unprotected eremomycin or vancomycin withRCH₂N(Fmoc)CH₂COOSu (where R=p-BuPh, p-CIPhPh, p-BuOPh and p-octylOPh)followed by deprotection with 10% diethylamine in DMSO. Eremomycinderivatives were obtained in 30-50% summary yields. Vancomycinderivatives were obtained in 30-60% summary yields. The starting glycinederivatives were synthesized as shown and described below in Scheme 1and the associated description of steps I, II and III.

I. Reductive Alkylation of Glycine (Synthesis of RCH₂NHCH₂COOH).

To a stirred solution of glycine (2 mmol) in THF:H₂O mixture (1:1) atroom temperature a solution of 1 mmol of an appropriate aldehyde in THFand 1.5 mmol of NaCNBH₃ were added portion-wise. The reaction mixturewas stirred for 4 h, then water was added. The resulting mixture wasevaporated under vacuum to remove THF and was extracted with petroleumether three times. Then the aqueous fraction was evaporated under vacuumwith silicagel to dryness and applied to a chromatographic column withsilicagel preequilibrated with CHCl₃. The column was eluted with aCHCl₃:MeOH:25% NH₄OH (3:1:0.05) system at a rate 10 mL/h, whilecollecting 5 mL fractions. The suitable fractions were combined andevaporated under vacuum to dryness. The yields were 30-50%.

II. Synthesis of RCH₂N(Fmoc)CH₂COOH

To a stirred solution of RCH₂NHCH₂COOH (1 mmol) in THF:H₂O mixture (1:1)at room temperature 3 mmol of triethylamine and a solution of 1.5 mmolof FmocOSu in THF were added portion-wise. The reaction mixture wasstirred for 4 h, then water was added. The resulting mixture wasevaporated under vacuum to remove THF and was extracted with petroleumether three times. Then the aqueous fraction was evaporated under vacuumwith silicagel to dryness and applied to a chromatographic column withsilicagel preequilibrated with CHCl₃. The column was eluted with aCHCl₃:MeOH:25% NH₄OH (5:1:0.05) system at a rate 10 mL/h, whilecollecting 5 mL fractions. The suitable fractions were combined andevaporated under vacuum to dryness. The yields were 50-80%.

III. Synthesis of RCH₂N(Fmoc)CH₂COOSu

To a stirred solution of RCH₂N(Fmoc)CH₂COOH (1 mmol) in CH₂Cl₂ at 0-5 ⁰C1.3 mmol of HOSu were added and a solution of 1.2 mmol of DCC in THF wasadded drop-wise. The reaction mixture was stirred for 4 hours, then aprecipitate of dicyclohexylurea was filtered off. The organic layer wasconcentrated under vacuum to a small volume and a precipitated solid ofdicyclohexylurea was filtered off again. The organic layer wasevaporated under vacuum to dryness.

Preparation of N′-(p-OctylOPhCH₂NHCH₂CO)vancomycin

P-OctylOPhCH₂N(Fmoc)CH₂COOSu was prepared as shown at the Scheme 1 in20% summary yield starting from glycine.

Then, to a stirred solution of 1800 mg (1.25 mmol) of vancomycin (base)in a 30 mL DMSO:H₂O (4:1) mixture, 0.16 mL (1.25 mmol) of Et₃N and 1165mg (1.9 mmol) of p-octylOPhCH₂N(Fmoc)CH₂COOSu were added. The reactionmixture was stirred at room temperature for 5 h, then 3 mL of Et₂NH wereadded. The reaction mixture was stirred at room temperature for 1 h,then it was added to 200 mL of acetone. A solid precipitate was filteredoff, washed with acetone and dried under vacuum. The resulting dryprecipitate was then dissolved in a H₂O:THF (1:1) mixture and evaporatedwith a small amount of silanized silicagel under vacuum. This solutionwas applied to a chromatographic column with silanized silicagel (3×120cm) preequilibrated with H₂O. The column was eluted firstly with H₂O(1000 mL) at a rate 10 mL/h, while collecting 5 mL fractions. Thefractions containing vancomycin were collected. The column was theneluted with 0.02 M CH3COOH (1000 mL) at a rate 10 mL/h, while collecting5 mL fractions. Then the column was eluted with 15% MeOH in 0.02 MCH3COOH (500 mL) at the same rate, and the fractions containing theproduct of the reaction were collected. Then the column was eluted with30% MeOH in 0.02 M CH3COOH (1000 mL) at the same rate, and the suitablefractions containing the product of the reaction were collected. All thesuitable fractions of N′-{p-octylOPhCH₂NHCH₂CO)vancomycin were combinedand concentrated under vacuum to a small volume (˜10 mL). Then 30 mL ofacetone were added and this mixture was added to 250 mL of Et₂O toprecipitate the product. A solid precipitate was filtered off, washedwith Et₂O, and dried under vacuum to give 904 mg (42%) ofN′-{p-octylOPhCH₂NHCH₂CO)vancomycin.

The purification of the eremomycin and vancomycin derivativessynthesized in this manner was performed using column chromatography onsilanized silica gel. The progress of the reactions, the components ofthe column eluates and the purity of the final compounds were checked byTLC in the systems: EtOAc-n-PrOH-25% NH₄OH 1:1:1 or 3:2:2 andn-BuOH—AcOH—H₂O 5:1:1. Additionally, the purity of the derivatives forin vivo study was controlled by HPLC.

Example 2 Preparation of N′-[C₈H₁₇OC₆H₄CH₂NHCH(CH₃)CO] Vancomycin

Cmpd R chemical formula MW 301 p-C₈H₁₇0BnNHCH(CH₃)CO C₈₄H₁₀₂N₁₀O₂₆Cl₂1736.6

I. Reductive Alkylation of L-alanine (Synthesis ofp-C₈H₁₇OC₆H₄CH₂NHCH(CH₃)COOH)

To a stirred solution of L-alanine (1 mmol) in THF-H₂O mixture (1:1) at26-28° C. a solution of 1 Mmol of p-C₈H₁₇OC₆H₄CHO in THF and 0.75 mmolof NaCNBH₃ were added portion-wise. The reaction mixture was stirred at26-28° C. for 4 h, then water was added. The resulting mixture wasevaporated under vacuum to remove THF and to precipitate the product ofthe reaction. The precipitate was filtered off and washed with icy coldwater. The solid was dissolved in THF-H₂O mixture (1:1) and this mixturewas kept at 5° C. for 18 h. A white solid (L-alanine) was filtered offand washed with icy cold water, the filtrate was evaporated under vacuumto remove THF and to precipitate the product of the reaction. Theprecipitate was washed with acetone and dried under vacuum. The yieldwas about 30-40%.

II. Synthesis of p-C₈H₁₇OC₆H₄CH₂NFmocCH₂(CH₃)COOH

To a stirred solution of N-(p-C₈H₁₇OC₆H₄CH₂)-(L)-alanine (1 mmol) inTHF-H₂O mixture (1:1) at room temperature 3 mmol of triethylamine and asolution of 1.5 mmol of FmocOSu in THF were added portion-wise. Thereaction mixture was stirred for 4 h, then water was added. Theresulting mixture was evaporated under vacuum to remove THF and wasextracted with petroleum ether. The organic layer was washed with water.The aqueous fractions were combined, evaporated under vacuum with silicagel to dryness and applied to a chromatographic column with silica gelpreequilibrated with CHCl₃. The column was eluted with a CHCl₃:MeOH:25%NH₄OH (5:1:0.05) system at a rate 10 mL/h, while collecting 5 mLfractions. The suitable fractions were combined and evaporated undervacuum to dryness. The yield was about 60-80%. III. Synthesis ofp-C₈H₁₇OC₆H₄CH₂NFmocCH(CH₃)COOSu

To a stirred solution of p-C₈H₁₇OC₆H₄CH₂NFmocCH(CH₃)COOH (1 mmol) inCH₂Cl₂ at 0-5 ⁰C 1.3 mmol of HOSu was added and, afterwards, a solutionof 1.2 mmol of DCC in THF drop-wise. The reaction mixture was stirredfor 4 h, then the precipitate of dicyclohexylurea was filtered off. Theorganic layer was concentrated under vacuum to a small volume and aprecipitated solid of dicyclohexylurea was filtered off again. Theorganic layer was evaporated under vacuum to dryness.P—C₈H₁₇OC₆H₄CH₂NFmocCH(CH₃)COOSu was obtained in summary yield of about20-30% starting from L-alanine according to Scheme 2.

Synthesis of N′-[C₈H₁₇OC₆H₄CH₂NHCH(CH₃)CO]Vancomycin

To a stirred solution of 360 mg (0.25 mmol) of vancomycin (base) in 7.5mL a DMSO-H₂O (4:1) mixture 32 μL (0.25 mmol) of triethylamine and 627mg (0.38 mmol) of starting amino acid derivativep-C₈H₁₇OC₆H₄CH₂NFmocCH(CH₃)COOSu were added. The reaction mixture wasstirred at room temperature for 5 h, then 0.75 mL of Et₂NH were added.The reaction mixture was stirred at room temperature for 1 h, then itwas added to 100 mL of acetone. A solid precipitate was filtered off,washed with acetone and dried under vacuum. Then it was dissolved in anH₂O-THF (1:1) mixture, evaporated with a small amount of silinizedsilica gel under vacuum and applied to a chromatographic column withsilinized silica gel (2×60 cm) preequilibrated with H₂O. The column waseluted firstly with H₂O (200 mL) at a rate 10 mL/h, while collecting 5mL fractions. The column was eluted with 0.02 M CH₃COOH (300 mL) at arate 10 mL/h, while collecting 5 mL fractions. The fractions containingvancomycin were collected. Then the column was eluted with 10% MeOH in0.02 M CH3COOH (250 mL) at the same rate followed with 20% MeOH in 0.02M CH₃COOH (250 mL) to elute side products. The fractions containing thedesired product were collected when the column was eluted with 40% MeOHin 0.02 M CH₃COOH. All the suitable fractions of the product werecombined and concentrated under vacuum to a small volume (˜2 mL). THF (2mL) was added then 20 mL of acetone were added to this mixture. Theresulting mixture was added to 80 mL of Et₂O to precipitate the reactionproduct. A solid precipitate was filtered off and washed with acetone,then dried in vacuum. The yield was 130 mg (30%).

Example 3 Synthesis of N′-Aminoacyl (Non-Glycyl) Derivatives ofEremomycin

N′-Acylated eremomycin derivatives substituted with amino acids orN-alkylated amino acids and a vancomycin derivative were prepared by thetreatment of an antibiotic with N-hydroxysuccinimide ester ofN-Fmoc-derivatives of amino acids or N-alkylated amino acids, followedby deprotection with 10% diethylamine in DMSO gave a desirable productin 10-50% summary yields. The starting derivatives of amino acids weresynthesized as shown and described below with reference to Scheme 3.

Preparation of N-Hydroxysuccinimide Ester of N^(α),N^(ε)-di-Fmoc-L-Lys(Scheme 3a)

N-Hydroxysuccinimide ester of N^(α),N^(ε)-di-Fmoc-L-Lys was obtainedfrom N^(α),N^(ε)-di-Fmoc-L-Lys by the method C (see below).

Preparation of N-hydroxysuccinimide Ester of N-Fmoc-L-Phe andN-Fmoc-D-Phe (scheme 3b).

N-Hydroxysuccinimide ester of N-Fmoc-L-Phe was obtained by the method Cfrom N-Fmoc-L-Phe, prepared from L-Phe by the method B.

Preparation of N-Hydroxysuccinimide Ester of N-Fmoc-(Bn-O-L-Tyr) (Scheme3b).

N-Fmoc-(Bn-O-L-Tyr) was obtained starting from Bn-O-L-Tyr by the methodB. N-hydroxysuccinimide ester of N-Fmoc-(Bn-O-L-Tyr) was prepared by themethod C.

Preparation of N-Hydroxysuccinimide Ester ofN^(α)—R—N^(α),N^(δ)-di-Fmoc-L-Orn (Scheme 3c).

N^(α)—R—N^(δ)-Boc-L-Om [R=p-(C₈H₁₇—O-Ph)—CH₂— or p-(BuPh)—CH₂—] wasobtained from N^(δ)-Boc-L-Om and R—CHO by the method A. ThenN^(α)—R—N^(δ)-Boc-L-Om was treated by TFA at room temperature for 30 minto give N^(α)—R-L-Orn. MeOH was added and the solution was evaporatedunder vacuum to dryness. This operation was repeated for 3 times.N^(α)—R—N^(α),N^(δ)-di-Fmoc-L-Om was obtained starting fromN^(α)—R-L-Orn as described in method B, but a solution of 2.2 eq. ofFmocCl in THF was added dropwise to a cooled (0-5° C.) solution of 1 eq.of N^(α)—R-L-Om and 5 eq. of K₂CO₃ in a THF-H₂O (1:1) mixture.N-hydroxysuccinimide ester of N^(α)—R—N^(α),N^(δ)-di-Fmoc-L-Orn wasprepared by the method C.

Method A. Reductive Alkylation

To a stirred solution of an N^(δ)-Boc-L-Orn (2 mmol) in THF:H₂O mixture(1:1) at room temperature a solution of 1 mmol of an appropriatealdehyde in THF and 1.5 mmol of NaCNBH₃ were added portion-wise. Thereaction mixture was stirred for 4 h, then water was added. Theresulting mixture was evaporated under vacuum to remove THF and wasextracted with petroleum ether. The aqueous fraction was evaporatedunder vacuum with silica gel to dryness and applied to a chromatographiccolumn with silica gel preequilibrated with CHCl₃. The column was elutedwith a CHCl₃:MeOH:25% NH₄OH (5:1:0.05) system at a rate 10 mL/h, whilecollecting 5 mL fractions. The suitable fractions were combined andevaporated under vacuum to dryness. The yields were 30-50%.

Method B. Preparation of N-Fmoc Derivatives

To a stirred solution of amino acid (1 mmol) in THF:H₂O mixture (1:1) atroom temperature 4 mmol of triethylamine and a solution of 1.5 mmol ofFmocOSu in THF were added portion-wise. The reaction mixture was stirredfor 4 h, then water was added. The resulting mixture was evaporatedunder vacuum to remove THF and was extracted with petroleum ether. Theaqueous fraction was evaporated under vacuum with silica gel to drynessand applied to a chromatographic column with silica gel preequilibratedwith CHCl₃. The column was eluted with a CHCl₃:MeOH:25% NH₄OH(5:1:0.05), according to Scheme 2b, or a (7:1:0.05), according to Scheme2c, system at a rate 10 mL/h, while collecting 5 mL fractions. Thesuitable fractions were combined and evaporated under vacuum to dryness.The yields were 50-80%.

Method C. Preparation of N-Hydroxysuccinimide Ester

To a stirred solution of starting N-Fmoc derivative (1 mmol) in CH₂Cl₂at 0-5 ⁰C 1.3 mmol of HOSu were added and a solution of 1.2 mmol of DCCin THF was added drop-wise. The reaction mixture was stirred for 4 h,then the precipitate of dicyclohexylurea was filtered off. The organiclayer was concentrated under vacuum to a small volume and a precipitatedsolid of dicyclohexylurea was filtered off again. The organic layer wasevaporated under vacuum to dryness.

Preparation of N′-Substituted Glycipeptide Derivatives

To a stirred solution of 0.5 mmol of an antibiotic (base) in 15 mLDMSO:H₂O (4:1) mixture 0.5 mmol of triethylamine and 0.75 mmol ofstarting amino acid derivative, prepared according to scheme 2, wereadded. The reaction mixture was stirred at room temperature for 5 h,then 1.5 mL of Et₂NH weas added. The reaction mixture was stirred atroom temperature for 1 h, then it was added to 100 mL of acetone. Asolid precipitated was filtered off, washed with acetone and dried undervacuum. Then it was dissolved in a H₂O:THF (1:1) mixture, evaporatedwith a small amount of silanized silica gel under vacuum and applied toa chromatographic column with silanized silica gel (3×120 cm)preequilibrated with H₂O. The column was eluted firstly with H₂O (400mL) at a rate 10 mL/h, while collecting 5 mL fractions. The column wasthen eluted with 0.02 M CH₃COOH (500 mL) at a rate 10 mL/h, whilecollecting 5 mL fractions. The fractions containing an antibiotic werecollected. Then the column was eluted with 10% MeOH in 0.02 M CH3COOH(500 mL) at the same rate, and the fractions containing the product ofthe reaction were collected. Then the column was eluted with 20% MeOH in0.02 M CH₃COOH (500 mL), then 30% MeOH in 0.02 M CH₃COOH at the samerate, and the suitable fractions containing the product of the reactionwere collected. All the suitable fractions of desirable product werecombined and concentrated under vacuum to a small volume (˜3 mL). Then50 mL of acetone were added to precipitate the productsN-hydroxysuccinimide ester of N^(α),N^(δ)-di-Fmoc-L-Lys andN-hydroxysuccinimide ester of N-Fmoc-L-Phe, N-Fmoc-D-Phe andN-Fmoc-(Bn-O-L-Tyr). For N-hydroxysuccinimide ester ofN^(α)—R—N^(α),N^(δ)-di-Fmoc-L-Om, 8 mL of acetone were added and thismixture was added to 100 mL of Et₂O to precipitate the product. A solidprecipitated was filterred off and washed with acetone or Et₂O, thendried under vacuum. The yields were 30-50% for N-hydroxysuccinimideester of N^(α),N^(ε)-di-Fmoc-L-Lys and N-hydroxysuccinimide ester ofN-Fmoc-L-Phe, N-Fmoc-D-Phe and N-Fmoc-(Bn-O-L-Tyr) and about 10% forN-hydroxysuccinimide ester of N^(α)—R—N^(α),N^(δ)-di-Fmoc-L-Om.

Example 4 Preparation of (Adamantylamino)Amides of GlycopeptideAntibiotics or their Derivatives

To a stirred solution of an antibiotic or its derivative (e.g., preparedas described in Example 3) (0.1 mmol) in DMSO (4 mL) 2-amino-adamantanor 1-amino-adamantane (0.5 mmol), Et₃N (1 mmol) and HBPyU[O-benzotriazol-1-yl-N,N,N′,N′-bis(tetramethylene)uroniumhexafluorophosphate] or PyBOP [benzotriazol-1-yloxy)-tris-(pyrrolidino)phosphonium-hexafluorophosphate] (0.2 mmol) were added at roomtemperature in three portions with stirring over 1 h. After 4 h acetone(100 mL) was added to give a solid, which was washed with acetone anddried under vacuum to give the corresponding amide in about 90% yield.

Example 5 Additional Amide Derivatization and Evaluation

The synthesis of amides of eremomycin was performed by the condensationof unprotected eremomycin with an appropriate amine in the presence ofPyBOP as a condensing agent, according to procedures described inMiroshnikova O. V., Printsevskaya S. S., Olsufyeva E. N., Pavlov A. Y.,Nilius A., Hensey-Rudloff D., Preobrazhenskaya M. N. J. Antibiot., 2000.V.53. P. 286-293, incorporated herein by reference for all purposes. Theyields of the amides depend on the nature of the amines and were 40-80%(e.g., the yield of Compound 79 was 40%, while Compound 90 was obtainedin 80% yield). Most of the starting amines were not availablecommercially and were prepared as shown in Scheme 4.

The aminomethylated derivatives of eremomycin were obtained by theinteraction of eremomycin with an amine and 37% aqueous formaldehyde atpH 9-9.5 according to procedures described in Pavlov A. Y., Lazhko E.I., Preobrazhenskaya M. N. J. Antibiot. 1996, V. 50, P. 509-513,incorporated herein by reference for all purposes. In contrast toeremomycin amides, the synthesis of the aminomethylated derivatives gavebetter yields for secondary amines than for primary amines (e.g.,Compound 72-40% and Compound 73-60%).

The amides of the aminomethylated derivatives were prepared by theamidation of the aminomethylated derivatives. The best results wereobtained by the amidation with the usage of an amine excess (˜5 times).The summary yields of the amides of the aminomethylated derivatives(starting from eremomycin) were 20-50%.

The amides of N-allyl-eremomycin and quaternary salt ofN,N-dimethyl-eremomycin were obtained by the reaction of the appropriateamide with allyl bromide or methyl iodide in DMSO in the presence ofNaHCO₃. In the case of allyl bromide the yield of the product was about60%, while methyl iodide gave 90% yield of the target amide ofN,N-dimethyl-eremomycin. Thus the summary yield of these derivatives was45-70%. The decyldimethylaminopropylamide of N,N-dimethyl-eremomycin(Compound 70 was prepared with 65% yield.

The purification of the derivatives of eremomycin was performed usingcolumn chromatography on CM-32-cellulose or silanized silica gel asdescribed in Pavlov A. Y., Berdnikova T. F., Olsufyeva E. N.,Miroshnikova O. V., Fillipposianz S. T., Preobrazhenskaya M. N., SottaniC., Colombo L., Goldstein B. P. J. Antibiot., 1996, V. 49, P. 194-198;and Miroshnikova O. V., Printsevskaya S. S., Olsufyeva E. N., Pavlov A.Y., Nilius A., Hensey-Rudloff D., Preobrazhenskaya M. N. J. Antibiot.,2000. V.53. P. 286-293, incorporated herein by reference for allpurposes. The progress of the reactions, the components of the columneluates and the purity of the final compounds were checked by TLC in thesystems: EtOAc-n-PrOH-25% NH₄OH 1:1:1 or 3:2:2 and n-BuOH—AcOH—H₂O5:1:1. Additionally, the purity of the most active derivatives wascontrolled by HPLC. The structures of the eremomycin derivatives wereconfirmed by ¹H NMR and by the methods of chemical degradation (acidhydrolysis yielding unmodified eremosamine and altered aglycon and alsoEdman's degradation that shows the presence of the unsubstitutedN-terminal amino acid), according to procedures described in thereferences noted above.

Example 6 Antibacterial Evaluation

Antibacterial activity in vitro was investigated by broth microdilutionmethod in Meuller-Hinton broth as recommended by NCCLS. All strainstested were clinical isolates either sensitive or resistant to naturalglycopeptides. Results are reported in the tables as MIC (minimalinhibitory concentration) in μg/ml. Most of the compounds synthesizedhave activity comparable with vancomycin against sensitive bacteria,Compound 70 being an exception. It is the most active derivative oferemomycin among all the compounds investigated against clinicalisolates of vancomycin sensitive gram-positive bacteria. All derivativesof eremomycin are more active than natural glycopeptides (Ere, Vanco,Teico) against GISA and GRE. Compounds 72, 87, 90 and 95 are the mostactive against GISA. Compounds 70, 72, 75, 76, 90 and 95 demonstratealso rather good activity against GRE strains (between 4-16 mcg/ml),however some lower than LY 333328.

Analysis of the MIC values obtained shows that the introduction of amoiety containing the quaternary fragment —N⁺R₁R₂C₁₀H₂₁ presents aproductive approach to the synthesis of derivatives with high activityagainst GISA and GRE. The positive influence of quartenization onantibacterial activity is clearly seen after comparison of MIC valuesfor Compound 89 with that for Compounds 90 or 95. The transformation ofthe group —NHC₁₀H₂₁ (89) into —N⁺Me₂C₁₀H₂₁ (90) or —N⁺Allyl₂C₁₀H₂₁ (95)leads to the increase of the activity up to 2-8 times against sensitiveand resistant bacteria. It is interesting also to note that Compounds 92and 96 containing two C₁₀H₂₁ moieties retain good activity against bothresistant and sensitive bacteria, while earlier it was concluded thatthe introduction of two hydrophobic non-quartenized substituents leadsto the significant decrease of antibacterial activity (more than by oneorder). The investigation of SAR for compounds containing the quaternaryfragment —N⁺R₁R₂C₁₀H₂₁ shows that the length of the spacer between thismoiety and the framework of eremomycin (Compounds 46 and 90) has nosignificant influence on the antibacterial activity. The nature(hydrophobicity) of the spacer (Compounds 88, 90 and 91) seems to bemore important.

Tables

The following tables identify specific species of compounds according tothe present invention and information concerning their associatedantibacterial activity. The glycopeptides were tested against a varietyof strains, indicated below, including Staphylococus epidermidis,Staphylococus haemolyticus, glycopeptide-intermediate Staphylococusaureus (GISA), glocopeptide-sensitive Enterococcus faecalis (GSE), andglocopeptide-resistant Enterococcus faecalis (GRE). Results are shown inthe table as minimum inhibitory concentration (MIC) in units of μg/ml:TABLE 1 N′-alkylglucyl- and N′-acylglycylsubstituted eremomycinderivatives of the formula:

wherein R_(A) is as indicated for each compound: Compound # R_(A) 44p-Cl-PhBn HNCH₂CO 194 (C₁₀H₂₁)₂HNCH₂CO 57 p-BuBnHNCH₂CO 187Bu₂NBnHNCH₂CO 293 p-F-BnHNCH₂CO 294 p-CF₃-BnHNCH₂CO 192 stilbenylHNCH₂CO287 (phenanthren-9-yl)CH₂HNCH₂CO 292 (fluoren-2-yl)CH₂HNCH₂CO 296(quinolin-2-yl)HNCH₂CO 193 p-BuOBnHNCH₂CO 214 p-C₈H₁₇OBnHNCH₂CO 223p-BnOBnHNCH₂CO 224 5-BnO-(indol-3-yl)CH₂HNCH₂CO 2251-Bn(indol-3-yl)CH₂HNCH₂CO 186 C₉H₁₉COHNCH₂CO 221 FmocHNCH₂CO 222AdocHNCH₂CO

TABLE 1a Antibacterial activity of N′-alkylglucyl- andN′-acylglycylsubstituted eremomycin derivatives Cmpd/ 533 3797 S. aureus3798 S. aureus 568 E. faecium 559 E. faecalis 569 E. faecium 560 E.faecalis Strain S. epidermidis 602 S. haemolyticus (GISA) (GISA) (GSE)(GSE) (GRE) (GRE) 44 0.5 2 4 4 1 1 8 8 194 2 2 8 8 2 4 8 16 57 0.5 2 4 40.5 0.5 4 4 187 1 1 8 8 1 2 32 32 293 4 4 8 8 4 4 16 16 294 1 1 4 4 0.50.5 >64 >64 192 0.5 0.5 8 8 0.5 1 4 8 287 2 2 16 16 2 2 >64 >64 292 4 48 8 2 2 32 32 296 4 4 >32 >32 2 2 >64 >64 193 0.5 1 4 4 1 2 64 64 214 11 4 4 0.5 1 8 8 223 2 4 8 8 2 2 16 64 224 2 2 8 8 1 2 16 >64 225 1 2 4 44 2 16 64 186 2 2 16 16 1 2 >64 >64 221 n.t n.t n.t n.t 0.5 0.5 64 >64222 n.t n.t n.t n.t 1 2 >64 >64

TABLE 2 N′-alkylglycylsubstituted vancomycin derivatives of the formula:

wherein R_(B) is as indicated for each compound: Compound # R_(B) 210p-Cl-PhBnHNCH₂CO 291 p-F-BnHNCH₂CO 290 p-CF₃-BnHNCH₂CO 182 p-BuBnHNCH₂CO218 p-BuOBnHNCH₂CO 220 p-C₈H₁₇OBnHNCH₂CO 298 (quinolin-2-yl)HNCH₂CO

TABLE 2a Antibacterial activity of N′-alkylglycylsubstituted vancomycinderivatives Cmpd/ 533 3797 S. aureus 3798 S. aureus 568 E. faecium 559E. faecalis 569 E. faecium 560 E. faecalis Strain S. epidermidis 602 S.haemolyticus (GISA) (GISA) (GSE) (GSE) (GRE) (GRE) 210 0.13 0.25 1 10.25 0.5 16 16 291 4 4 4 4 1 1 8 8 290 4 4 4 4 4 2 >64 >64 182 0.25 0.52 2 0.5 1 32 32 218 0.13 0.13 0.5 1 0.5 1 >64 >64 220 0.5 1 2 2 0.250.25 2 4 298 8 8 8 8 4 2 >64 64CHIRP020

TABLE 3 Eremomycin derivatives N′-substituted by non-glycine amino acidshaving the formula:

wherein R_(C) is as indicated for each compound: Compound # antibioticR_(C) 229 eremomycin D-Phe 230 eremomycin L-Phe 228 eremomycinBn-O-L-Tyr 203 eremomycin Lys 242 eremomycin N^(α)-p-BuBn-L-Orn (analogof #57) 241 vancomycin N^(α)-p-C₈H₁₇-O-Bn-L-Orn (analog of #220)

TABLE 3a Antibacterial activity of Eremomycin derivatives N′-substitutedby non- glycine amino acids Cmpd/ 533 3797 S. aureus 3798 S. aureus 568E. faecium 559 E. faecalis 569 E. faecium 560 E. faecalis Strain S.epidermidis 602 S. haemolyticus (GISA) (GISA) (GSE) (GSE) (GRE) (GRE)229 1 1 >32 >32 0.25 0.5 >64 >64 230 4 4 >32 >32 2 2 >64 >64 228 4 4 1616 2 2 >64 >64 203 0.13 0.13 4 8 0.25 0.25 16 >64 242 0.25 1 4 4 0.5 1 88 241 0.5 1 2 2 1 1 16 16

TABLE 4 Double modified eremomycin derivatives of the formula:

wherein R_(D) and R_(E) are as indicated for each compound: Compound #R_(D) R_(E) 77 p-BuBnHNCH₂CO CH₃NH 263 p-BuBnHNCH₂CO (Adam-2)NH 264p-BuBnHNCH₂CO (Adam-1)CH(CH₃)N 265 p-C₈H₁₇-OBnHNCH₂CO (Adam-2)NH 266p-C₈H₁₇-OBnHNCH₂CO (Adam-1)CH(CH₃)N 275 p-Cl-PhBnHNCH₂CO p-F-BnNH 213 H(Adam-2)NH 262 H (Adam-1)CH(CH₃)N

TABLE 4a Antibacterial activity of double modified eremomycinderivatives Cmpd/ 533 3797 S. aureus 3798 S. aureus 568 E. faecium 559E. faecalis 569 E. faecium 560 E. faecalis Strain S. epidermidis 602 S.haemolyticus (GISA) (GISA) (GSE) (GSE) (GRE) (GRE) 77 1 2 2 2 2 2 8 8263 8 8 16 16 8 8 8 8 264 4 8 8 16 4 4 8 8 265 32 32 >32 >32 n.t n.t n.tn.t 266 16 32 >32 >32 n.t n.t n.t n.t 213 0.25 0.25 1 2 0.5 0.5 4 8 2620.5 1 4 2 0.5 1 16 16

TABLE 5 Double modified vancomycin derivatives of the formula:

wherein R_(F) and R_(G) are as indicated for each compound: Compound #R_(F) R_(G) 276 p-BuBnHNCH₂CO p-F-BnNH 277 p-C₈H₁₇-OBnHNCH₂CO p-F-BnNH288 H p-F-BnNH

TABLE 5a Antibacterial activity of double modified vancomycinderivatives GINA/ 533 3797 S. aureus 3798 S. aureus 568 E. faecium 559E. faecalis 569 E. faecium 560 E. faecalis Strain S. epidermidis 602 S.haemolyticus (GISA) (GISA) (GSE) (GSE) (GRE) (GRE) 276 0.5 2 2 2 2 4 1616 277 4 8 8 4 4 4 8 8 288 2 1 4 4 0.13 0.13 >64 >64

CONCLUSION

Although the foregoing invention has been described in some detail forpurposes of clarity of understanding, it will be apparent that certainchanges and modifications may be practiced within the scope of theappended claims. It should be noted that there are many alternative waysof implementing both the processes and compositions of the presentinvention. Accordingly, the present embodiments are to be considered asillustrative and not restrictive, and the invention is not to be limitedto the details given herein, but may be modified within the scope andequivalents of the appended claims.

1. A compound having a formula selected from the group consisting of:

wherein, R₁ is C(═O)CR₇R_(7a)NR₈R_(8a), wherein, R₇ and R_(7a), areindependently hydrogen, the side chain of a naturally occurring ornon-naturally occurring amino acid, alkyl, or alkyl substituted with oneor more substituents selected from the group consisting of halogen,hydroxy, alkoxy, alkoxyalkoxy, carboxyl, carboxyl ester,—C(═O)NR₈R_(8a), —NR₈R_(8a), aryl, substituted aryl, heteroaryl,substituted heteroaryl, mercapto, or thioalkoxy, or R₇ and R_(7a)together with the atom to which they are attached form a cycloalkyl ringwhich optionally contains a heteroatom selected from the groupconsisting of optionally substituted O, N, and S; R₈ and R_(8a) areindependently selected from the group consisting of hydrogen andunsubstituted or substituted alkyl, alkenyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, arylalkyl, alkylaryl, and heteroaryl, said aryl,alkylaryl, arylalkyl or heteroaryl group optionally containing one ormore optionally substituted aryl, heteroaryl, or condensed rings, or R₈and R_(8a) together with the atom to which they are attached form acycloalkyl ring which optionally contains a heteroatom selected from thegroup consisting of optionally substituted O, N, and S; R_(1A) isselected from the group consisting of H, CHR₅R_(5a), and C(═O)₆,wherein, R₅ and R_(5a) are independently selected from the groupconsisting of hydrogen and unsubstituted or substituted alkyl, alkenyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, arylalkyl, alkylaryl,and heteroaryl, said aryl, alkylaryl, arylalkyl or heteroaryl groupoptionally containing one or more optionally substituted aryl,heteroaryl, or condensed rings, or R₅ and R_(5a) together with the atomto which they are attached form a cycloalkyl ring which optionallycontains a heteroatom selected from the group consisting of optionallysubstituted O, N, and S, and R₆ is selected from the group consisting ofunsubstituted or substituted alkyl, alkenyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl, arylalkyl, alkylaryl, and heteroaryl containinga heteroatom selected from the group consisting of optionallysubstituted O, N, and S, said aryl, alkylaryl, arylalkyl, or heteroarylgroup optionally containing one or more optionally substituted aryl,heteroaryl, or condensed rings; R₂ is selected from the group consistingof, (1) OH, (2) 1-adamantanamino, (3) 2-adamantanamino, (4)3-amino-1-adamantanamino, (5) 1-amino-3-adamantanamino, (6)3-loweralkylamino-1-adamantanamino, (7)1-loweralkylamino-3-adamantanamino, (8) amino, (9) NR₉R_(9a) wherein R₉and R_(9a) are independently selected from the group consisting ofhydrogen, loweralkyl or substituted loweralkyl, or  R₉ and R_(9a)together with the atom to which they are attached form a 3-10 memberedheterocycloalkyl ring, which may optionally be substituted with one ormore substituents independently selected from the group consisting of(a) halogen, (b) hydroxy, (c) C₁-C₃-alkoxy, (d)C₁-C₃-alkoxy-C₁-C₃-alkoxy, (e) oxo, (f) C₁-C₃-alkyl, (g)halo-C₁-C₃-alkyl, and (h) C₁-C₃-alkoxy-C₁-C₃-alkyl; R_(2A) is selectedfrom the group consisting of (1) 1-adamantanamino, (2) 2-adamantanamino,(3) 3-amino-1-adamantanamino, (4) 1-amino-3-adamantanamino, (5)3-loweralkylamino-1-adamantanamino, (6)1-loweralkylamino-3-adamantanamino; and R₃ is selected from the groupconsisting of hydrogen and aminoloweralkyl, wherein the aminoloweralkylamino group is further substituted with unsubstituted or substitutedalkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, arylalkyl, alkylaryl,alkoxy, aryloxy, substituted alkoxy, and substituted aryloxy; or apharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomeror prodrug thereof.
 2. The compound of claim 1, wherein R₅ is hydrogenand R_(5a) is selected from the group consisting of unsubstituted orsubstituted alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,aryl, arylalkyl, alkylaryl, and heteroaryl, said aryl, alkylaryl,arylalkyl or heteroaryl group optionally containing one or moreoptionally substituted aryl, heteroaryl, or condensed rings, or R₅ andR_(5a) together with the atom to which they are attached form acycloalkyl ring which optionally contains a heteroatom selected from thegroup consisting of optionally substituted O, N, and S.
 3. The compoundN′-p-BuBnHNCH₂CO eremomycin.
 4. The compound N′-stilbenzylHNCH₂COeremomycin.
 5. The compound N′-p-C₈H₁₇OBnHNCH₂CO vancomycin.
 6. Thecompound N′-p-C₈H₁₇OBnHNCH(CH₃)CO vancomycin
 7. The compound2-adamantanamino eremomycin.
 8. The compound of claim 1, wherein R₆ is aβ-amino acid analog comprising a —CH₂CHNH— portion.
 9. The compound ofclaim 8, wherein R₆ is selected from the group consisting ofCH₂C(R₇)(R_(7a))(NR₈R_(8a)) wherein R₇, R_(7a), R₈, and R_(8a) arepreviously defined or —CR₇R_(7a) together with NR₈R_(8a) form apyrrolidine ring.
 10. The compound of claim 1, whereinC(═O)CR₇R_(7a)NR₈R_(8a) is selected from the group consisting of aminoacid moieties.
 11. The compound of claim 10, wherein R₇, R₈ and R_(8a)are each H and R_(7a) is selected from the group consisting of H, CH₃,CH(CH₃)₂, CH₂CH(CH₃)₂, CH(CH₃)CH₂CH₃, (CH₂)₄NH₂, CH₂OH, CH(OH)CH₃,CH₂COOH, (CH₂)₂COOH, CH₂C(═O)NH₂, (CH₂)₂C(═O)NH₂, CH₂SH, (CH₂)₂SCH₃,(CH₂)₃NHC(═NH)NH₂, CH₂C₆H₅, CH₂C₆H₄OH, CH₂(4-imidazoyl) andCH₂(3-indolyl), or —CR₇R_(7a) together with NR₈R_(8a) form a pyrrolidinering.
 12. The compound of claim 1, wherein R₇ is H and R_(7a) isselected from the group consisting of (1) hydrogen, (2) C₁-C₁₂-alkyl,and (3) C₁-C₁₂-alkyl substituted with one or more substituents selectedfrom the group consisting of (a) halogen, (b) hydroxy, (c) C₁-C₃-alkoxy,(d) C₁-C₃-alkoxy-C₁-C₃-alkoxy, (e) —CO₂R₅ wherein R₅ is hydrogen,loweralkyl or substituted loweralkyl, (f)—C(═O)NR₉R_(9a), (g) amino, and(h) —NR₉R_(9a), or  R₉ and R_(9a) together with the atom to which theyare attached form a 3-10 membered heterocycloalkyl ring optionallysubstituted with one or more substituents independently selected fromthe group consisting of (i) halogen. (ii) hydroxy, (iii) C₁-C₃-alkoxy,(iv) C₁-C₃-alkoxy-C₁-C₃-alkoxy, (v) oxo, (vi) C₁-C₃-alkyl, (vii)halo-C₁-C₃-alkyl, and (viii) C₁-C₃-alkoxy-C₁-C₃-alkyl, (i) aryl, (j)substituted aryl, (k) heteroaryl, (l) substituted heteroaryl, (m)mercapto, and (n) C₁-C₃-thioalkoxy.
 13. The compound of claim 1, whereinR₈ and R_(8a) are independently selected from the group consisting of,(1) hydrogen, (2) C₁-C₁₂-alkyl, (3) C₂-C₁₂-alkyl substituted with one ormore substituents selected from the group consisting of (a) halogen, (b)hydroxy, (c) C₁-C₃-alkoxy, (d) C₁-C₃-alkoxy-C₁-C₃-alkoxy, (e) amino, and(f) C₁-C₃-alkylamino, (4) C₁-C₁₂-alkyl substituted with aryl, (5)C₁-C₁₂-alkyl substituted with substituted aryl, (6) C₁-C₁₂-alkylsubstituted with heteroaryl, and (7) C₁-C₁₂-alkyl substituted withsubstituted heteroaryl; or R₈ and R_(8a) together with the atom to whichthey are attached form a C₃-C₇-heterocycloalkyl ring which, when thering is a 5- to 7-membered ring, optionally contains a hetero functionselected from the group consisting of —O—, —NH, —N(C₁-C₆-alkyl-)-,—N(aryl)-, —N(aryl-C₁-C₆-alkyl-)-, —N (substituted-aryl-C₁-C₆-alkyl-)-,—N(heteroaryl)-, —N(heteroaryl-C₁-C₆-alkyl-)-,—N(substituted-heteroaryl-C₁-C₆-alkyl-)-, and —S— or S(═O)_(n)— whereinn is 1 or
 2. 14. A pharmaceutical composition comprising atherapeutically effective amount of a compound of claim 1, together witha pharmaceutically acceptable carrier.
 15. A method of treating a mammalin need of such treatment comprising administering to the mammal anantibacterially effective amount of a compound of claim 1 together witha pharmaceutically acceptable carrier.
 16. A method of making a compoundof claim 1, comprising: modifying a glycopeptide scaffold selected formthe group consisting of eremomycin, A82846B, vancomycin, teicoplanin andA-40,926 scaffolds,

by a technique selected from the group consisting of, (a) acylation ofthe amino substituent on the amino-substituted sugar moiety of thecompound with an acyl group having the structure,—C(═O)CR₇R_(7a)NR₈R_(8a), (b) conversion of the acid moiety on themacrocyclic ring of the compound with a substituted amide as defined byR₂, and (c) a combination of (a) and (b) (d) a combination of (b) andacylation of the amino substituent on the amino-substituted sugar moietyof the compound with an acyl group having the structure,—C(═O)R₆, (e) a combination of (b) and alkylation of the aminosubstituent on the amino-substituted sugar moiety of the compound withan alkyl group having the structure,CHR₅R_(5a), to form a compound having a formula selected from the groupconsisting of:

wherein R₁, R_(1A), R₂, R_(2A), R₃, R₅, R_(5a), R₆, R₇, R_(7a), R₈, andR_(8a) are as defined herein.